title: Metagenomics for NoV surveillance: Outbreak-related sample sequencing
description: For Norovirus (NoV) genomic surveillance, metagenomics sequencing of viral nucleotides captured by VirCapSeq-VERT probe set, was performed. This metagenomics approach, in theory agnostic, offers the great advantage of unbiased detection of NoV, including the detection of novel NoV recombinants. Samples come from three different sequencing sets - Baseline Survey (BLS) samples, Outbreak-related (OB) samples, and two synthetic Test Set samples (TSS1 and TSS2). The samples from norovirus outbreaks associated with oysters included the clinical human stool samples from ill consumers and samples from the oyster batch linked to the outbreak were selected. A clarified 10% (w/v) fecal suspension was prepared in phosphate-buffered saline, centrifuged and the supernatant was filtered through a 0.45 -Âµm filter (Millipore). The viral-particle-containing filtrates were digested with a mixture of DNases and RNases (Strubbia et al. 2020). Viral RNA was extracted using the High Pure RNA isolation kit (Roche). Since stool samples are patient-derived, human data was removed by mapping against the human reference genome prior to submission.